MiR-96 promotes apoptosis of nucleus pulpous cells by targeting FRS2
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RESEARCH ARTICLE
MiR‑96 promotes apoptosis of nucleus pulpous cells by targeting FRS2 Xiaoxia Yang1 · Hengping Liu1 · Qingfeng Zhang1 · Kan Liu1 · Dong Yu1 · Yi Zhang1 · Zongting Shi1 Received: 5 March 2020 / Accepted: 5 June 2020 © Japan Human Cell Society 2020
Abstract This study aimed to investigate the molecular mechanism by which microRNA (miR)-96 regulates the progression of intervertebral disc degeneration (IDD). The expression of miR-96 in normal intervertebral discs and in IDD was detected by performing reverse transcription-quantitative PCR. CCK8 assay was applied to examine the proliferation of nucleus pulpous (NP) cells and flow cytometry was used to evaluate the cell apoptosis and cell cycle profile. In addition, the immunofluorescence analysis was employed to detect cell proliferation. The expressions of proteins were assessed by western blot analysis. TargetScan and miRDB were used to predict the target genes of miR-96. The results indicated that miR-96 expression was upregulated in IDD compared with normal intervertebral discs. Overexpression of miR-96 could significantly inhibit the proliferation of NP cells via inducing apoptosis and G1 arrest. In addition, fibroblast growth factor receptor substrate 2 (FRS2) was identified as the target of miR-96 and overexpression of FRS2 could revere the effect of miR-96 mimics in NP cells. Therefore, these findings demonstrated that miR-96 plays a critical role during the progression of IDD and miR-96 may serve as a target for the treatment of IDD. Keywords Intervertebral disc degeneration · miR-96 · Fibroblast growth factor receptor substrate 2 · Cell cycle
Introduction Intervertebral disc degeneration (IDD) is the leading cause of low back pain in the elderly, which was considered as a great burden on the public health [1–3]. Although significant advances have been made in identifying new therapeutic methods for IDD, the development of an effective therapy for this disease is very limited [2]. Therefore, it is urgent to investigate the molecular mechanisms of IDD to identify therapeutic targets for this condition. It is well known that the intervertebral disc comprises the nucleus pulpous (NP) and the annulus fibrosus, which are important for determining the maximum load of the spine [4]. It has been shown that NP is abundant in the extracellular matrix (ECM) of proteins; the deficiency of NP cells and degradation of ECM are responsible for the onset and progression of IDD [5]. Therefore, reduction of the NP cell death during the
* Xiaoxia Yang [email protected] 1
Department of Spine, Beijing University of Chinese Medicine Third Affiliated Hospital, No.51 Xiaoguan Street, Outside, Andingmen, Beijing 100029, China
progression of IDD may be an effective method to attenuate disc degeneration. MicroRNAs (miRNAs) are a class of non-coding RNAs with a length
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